Search

CN-121975680-A - Tobacco leaf extract degrading bacterium YY-2-4-1, screening method and application thereof, and method for treating tobacco leaf leaching wastewater by using same

CN121975680ACN 121975680 ACN121975680 ACN 121975680ACN-121975680-A

Abstract

The invention discloses a tobacco leaf extract degrading bacterium YY-2-4-1, a screening method and application thereof, and a method for treating tobacco leaf leaching wastewater by using the same, wherein the tobacco leaf extract degrading bacterium YY-2-4-1 is acidophilic oligoaeromonas and is preserved in China general microbiological culture collection center (CGMCC) with the preservation number of No.36759, according to the invention, the original strain obtained from a wastewater treatment plant for extracting wastewater from tobacco leaves is prepared from the tobacco leaf extract serving as a raw material through the steps of training, separating, purifying and the like of a culture medium containing single pollutant of the tobacco leaf extract, so that the pure strain YY2-4-1 is obtained, the degradation efficiency of the wastewater from the tobacco leaf extraction is 76.6%, the stable and efficient removal of the pollutant in the wastewater from the tobacco leaf extraction can be realized, and the wastewater treatment cost is reduced.

Inventors

  • LIU YU
  • LIU CHENG
  • LI YING

Assignees

  • 安徽焦甜香生物科技有限公司

Dates

Publication Date
20260505
Application Date
20260108

Claims (10)

  1. 1. The tobacco leaf extract degrading bacterium YY-2-4-1 is characterized in that the tobacco leaf extract degrading bacterium YY-2-4-1 is acidophilic oligoaeromonas and is preserved in China general microbiological culture collection center (CGMCC) No.36759.
  2. 2. A screening method of tobacco extract degrading bacteria YY-2-4-1 according to claim 1, characterized in that the screening method comprises the steps of: (1) Training strains, namely inoculating tobacco leaf leaching wastewater into an inorganic salt liquid culture medium containing tobacco leaf extracts for 3 days for 1 period, and gradually increasing COD concentration values of the culture medium to be 1500mg/L, 2000mg/L and 2500mg/L in sequence; (2) Strain screening, namely diluting the trained bacterial liquid by 10 -5 times, coating the bacterial liquid in an inorganic salt solid culture medium containing tobacco leaf extracts, and culturing at constant temperature for 60-72 hours; (3) Inoculating the strain cultured in the step (2) into an inorganic salt liquid culture medium containing tobacco leaf extracts, and carrying out constant-temperature vibration culture for 60-72 h; (4) Bacterial separation, namely diluting the bacterial liquid of the expanded culture by 10 -5 times, inoculating the bacterial liquid into an inorganic salt solid culture medium containing tobacco leaf extracts, and culturing at constant temperature for 60-72 h; (5) Strain purification, namely streaking and inoculating the single colony which grows fast in the step (4) into an LB solid culture medium, and culturing for 45-48 hours at constant temperature; (6) Repeating the steps (3), 4 and 5) until colony morphology on the LB solid medium is consistent and no second mixed bacteria exist, and marking the obtained strain as YY-2; (7) Strain re-screening, namely inoculating the strain YY-2 into an inorganic salt liquid culture medium containing tobacco leaf extracts for 3-5 days for 1 period, and gradually increasing the COD concentration value of the culture medium to 3500mg/L and 5000mg/L; (8) Repeating the steps (2), 3,4 and 5) to obtain the tobacco leaf extract degrading bacteria YY-2-4-1.
  3. 3. The screening method according to claim 2, wherein the pH value of the inorganic salt liquid medium containing the tobacco leaf extract is 7.5.+ -. 0.3, the composition is ：Na 2 HPO 4 ·12H 2 O 1.5-1.6g/L,KH 2 PO 4 0.3- 0.4g/L,MgSO 4 ·7H 2 O 0.1-0.15g/L,CaCl 2 0.05-0.1g/L, trace metal element solution 1mL/L, the composition of the tobacco leaf extract is 1.2g/L, and the composition of the trace metal element solution is ：FeSO 4· 7H 2 O 0.025g/L,ZnSO 4· 7H 2 O 0.01g/L,MnCl 2· 4H 2 O 0.03g/L,H 3 BO 4 0.3g/L,CoCL 2· 6H 2 O 0.2g/L,CuCI 2· 2H 2 O 0.01g/L,NiCl 2· 6H 2 O 0.02g/L,Na 2 MoO 4· 2H 2 O 0.03g/L.
  4. 4. The method according to claim 2, wherein the inorganic salt solid medium containing the tobacco leaf extract comprises adding 20g/L agar powder to the inorganic salt liquid medium containing the tobacco leaf extract.
  5. 5. The method according to claim 2, wherein the LB solid medium comprises 10g/L of tryptone, 5g/L of yeast extract, 10g/L of sodium chloride and pH 7.5.+ -. 0.3.
  6. 6. The screening method according to claim 2, wherein the temperature of the constant temperature culture and the constant temperature vibration culture is 35+ -3 ℃, and the rotation speed of the constant temperature vibration culture is 150-170 r/min.
  7. 7. The screening method according to claim 2, wherein in the step (1), the amount of the inoculated original bacterial liquid is 1-5% of the mass of the culture medium, in the step (2), the amount of inoculated bacterial liquid is 0.01-0.05 mL, and in the step (7), the amount of inoculated bacterial liquid is 1-5% of the mass of the culture medium.
  8. 8. The screening method according to claim 2, wherein in the step (1), the tobacco leaf leaching wastewater is wastewater generated by leaching the tobacco leaf extract, and the tobacco leaf extract is an extract or a tincture obtained by crushing, ethanol extraction, sedimentation, centrifugation, concentration, alcohol sedimentation, secondary centrifugation concentration and drying of tobacco leaves.
  9. 9. Use of the tobacco extract degrading bacterium YY-2-4-1 according to claim 1 in treating tobacco leaching wastewater.
  10. 10. A method for treating tobacco leaching wastewater by using the tobacco extract degrading bacterium YY-2-4-1 according to claim 1, characterized in that the method comprises the following steps: 1) Inoculating tobacco leaf extract degrading bacteria YY-2-4-1 into an LB liquid culture medium, and performing constant-temperature vibration culture for 20-24 hours to obtain seed liquid; 2) Inoculating the seed liquid into an LB liquid culture medium, culturing for 72 hours, pumping into an SBR biochemical tank, wherein the pH of tobacco leaf leaching wastewater fed into the SBR biochemical tank is 7.5+/-0.3, the COD is 2500-3500 mg/L, the temperature of the biochemical tank is controlled to be 32-38 ℃, the pH is controlled to be 7-8, the dissolved oxygen is controlled to be 2-4 mg/L, the MLSS is controlled to be 2000-4000mg/L, the volume load is controlled to be 0.1-0.3kg COD/(m 3 . D), and the residence time is controlled to be 2-5d.

Description

Tobacco leaf extract degrading bacterium YY-2-4-1, screening method and application thereof, and method for treating tobacco leaf leaching wastewater by using same Technical Field The invention belongs to the technical field of microbial engineering and environmental protection, and particularly relates to a tobacco leaf extract degrading bacterium YY-2-4-1, a screening method and application thereof, and a method for treating tobacco leaf leaching wastewater by using the same. Background The tobacco leaves contain components such as nicotine, solanesol, chlorogenic acid and the like, and the aroma components of the tobacco leaves are complex and classified according to different aroma-causing groups, and can be classified into acids, alcohols, ketones, aldehydes, esters, lactones, phenols, nitrogen heterocycles, furans, amides, ethers and hydrocarbons. The tobacco extract is a product which takes tobacco as a raw material and extracts specific required components through extraction, distillation and other modes. Tobacco extracts are often used to complement the tobacco flavor, enrich the leaf group formulation, and make the smoke richer and fuller. In the electronic atomized liquid product with the tobacco flavor, the tobacco extract is used as a main tobacco aroma substance, so that the tobacco aroma is reduced to a great extent. The tobacco leaching wastewater is high COD and high chroma organic wastewater generated in the tobacco leaching process, contains nicotine, protein and polyphenol substance residues, and is difficult to treat due to complex components, the biological treatment cannot reach the standard, and the chemical oxidation method has excessive cost. Along with the expansion of the market of electronic cigarettes, the amount of waste water extracted from tobacco leaves is increased, and the waste water cannot be discharged at will due to the pollution of the electronic cigarettes, so that the general treatment method is difficult and high in cost, and the finding of a proper treatment method is a problem which is needed to be solved currently. Disclosure of Invention In order to solve the technical problems, the invention provides the tobacco extract degrading bacterium YY-2-4-1, and the screening method and application thereof, wherein the tobacco extract degrading bacterium YY-2-4-1 can realize stable and efficient removal of pollutants in tobacco leaching wastewater and reduce wastewater treatment cost. The invention also provides a method for treating the tobacco leaching wastewater by utilizing the tobacco extract degrading bacteria YY-2-4-1, wherein the tobacco leaching wastewater is treated by utilizing the tobacco extract degrading bacteria YY-2-4-1 in an SBR biochemical tank, and after the tobacco extract degrading bacteria YY-2-4-1 are treated, the COD of precipitated effluent of the tobacco leaching wastewater is finally stabilized below 430mg/L, ammonia nitrogen is maintained below 13mg/L, and the requirement that the COD of effluent is lower than 500mg/L is met. In order to achieve the above purpose, the technical scheme adopted by the invention is as follows: The invention provides a tobacco leaf extract degrading bacterium YY-2-4-1, wherein the tobacco leaf extract degrading bacterium YY-2-4-1 is acidophilic oligomonad Stenotrophomonas acidaminiphila and is preserved in China general microbiological culture Collection center (CGMCC) No.36759, the preservation address is North Star Xway No. 1,3 in the Beijing Kogyo area, and the preservation time is 2025 years 11 months 25. The invention also provides a screening method of the tobacco leaf extract degrading bacteria YY-2-4-1, which comprises the following steps: (1) Training strains, namely inoculating tobacco leaf leaching wastewater into an inorganic salt liquid culture medium containing tobacco leaf extracts for 3 days for 1 period, and gradually increasing COD concentration values of the culture medium to be 1500mg/L, 2000mg/L and 2500mg/L in sequence; (2) Strain screening, namely diluting the trained bacterial liquid by 10 -5 times, coating the bacterial liquid in an inorganic salt solid culture medium containing tobacco leaf extracts, and culturing at constant temperature for 60-72 hours; (3) Inoculating the strain cultured in the step (2) into an inorganic salt liquid culture medium containing tobacco leaf extracts, and carrying out constant-temperature vibration culture for 60-72 h; (4) Bacterial separation, namely diluting the bacterial liquid of the expanded culture by 10 -5 times, inoculating the bacterial liquid into an inorganic salt solid culture medium containing tobacco leaf extracts, and culturing at constant temperature for 60-72 h; (5) Strain purification, namely streaking and inoculating the single colony which grows fast in the step (4) into an LB solid culture medium, and culturing for 45-48 hours at constant temperature; (6) Repeating the steps (3), 4 and 5) until colony morphology on the LB solid mediu